Benzodiazepine derivatives as inhibitors of gamma secretase

ABSTRACT

Compounds of formula (I) are disclosed. The compounds inhibit the action of gamma secretase, and hence find use in the treatment and prevention of Alzheimer&#39;s disease.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a U.S. National Phase application under 35 U.S.C. §371 of PCT Application No. PCT/GB01/04474, filed Oct. 8, 2001, whichclaims priority under 35 U.S.C. § 119 from GB Application No. 0025173.6,filed Oct. 13, 2000.

The present invention relates to a novel class of compounds, theirsalts, pharmaceutical compositions comprising them, processes for makingthem and their use in therapy of the human body. In particular, theinvention relates to compounds which modulate the processing of APP byγ-secretase, and hence are useful in the treatment or prevention ofAlzheimer's disease.

Alzheimer's disease (AD) is the most prevalent form of dementia.Although primarily a disease of the elderly, affecting up to 10% of thepopulation over the age of 65, AD also affects significant numbers of,younger patients with a genetic predisposition. It is aneurodegenerative disorder, clinically characterized by progressive lossof memory and cognitive function, and pathologically characterized bythe deposition of extracellular proteinaceous plaques in the corticaland associative brain regions of sufferers. These plaques mainlycomprise fibrillar aggregates of β-amyloid peptide (Aβ), and althoughthe exact role of the plaques in the onset and progress of AD is notfully understood, it is generally accepted that suppressing orattenuating the secretion of Aβ is a likely means of alleviating orpreventing the condition. (See, for example, ID research alert 19961(2):1–7; ID research alert 1997 2(1):1–8; Current Opinion in CPNSInvestigational Drugs 1999 1(3):327–332; and Chemistry in Britain,January 2000, 28–31.)

Aβ is a peptide comprising 39–43 amino acid residues, formed byproteolysis of the much larger amyloid precursor protein. The amyloidprecursor protein (APP or AβPP) has a receptor-like structure with alarge ectodomain, a membrane spanning region and a short cytoplasmictail. Different isoforms of APP result from the alternative splicing ofthree exons in a single gene and have 695, 751 and 770 amino acidsrespectively.

The Aβ domain encompasses parts of both extra-cellular and transmembranedomains of APP, thus its release implies the existence of two distinctproteolytic events to generate its NH₂— and COOH-termini. At least twosecretory mechanisms exist which release APP from the membrane andgenerate the soluble, COOH-truncated forms of APP (APP_(s)). Proteaseswhich release APP and its fragments from the membrane are termed“secretases”. Most APP_(s) is released by a putative α-secretase whichcleaves within the Aβ domain (between residues Lys¹⁶ and Leu¹⁷) torelease α-APP_(s) and precludes the release of intact Aβ. A minorportion of APP_(s) is released by a β-secretase, which cleaves near theNH₂-terminus of Aβ and produces COOH-terminal fragments (CTFs) whichcontain the whole Aβ domain. Finding these fragments in theextracellular compartment suggests that another proteolytic activity(γ-secretase) exists under normal conditions which can generate theCOOH-terminus of Aβ.

It is believed that at γ-secretase itself depends for its activity onthe presence of presenilin-1. In a manner that is not fully understood,full length presenilin-1 undergoes cleavage to a C-terminal fragment andan N-terminal fragment.

There are relatively few reports in the literature of compounds withinhibitory activity towards β- or γ-secretase, as measured in cell-basedassays. These are reviewed in the articles referenced above. Many of therelevant compounds are peptides or peptide derivatives.

WO95/14471 and WO95/14676 disclose classes of 3-acylaminobenzodiazepineswhich are antiarrhythmic agents, but do not disclose inhibition ofγ-secretase.

WO98/28268 discloses a broad range of compounds as inhibitors ofγ-secretase, including certain 3-acylamino-5-aryl-1,4-benzodiazepines,but there is no disclosure of compounds in accordance with the presentinvention.

According to the invention, there is a provided a compound of formula I:

wherein:

n is 0–3;

each R^(x) independently represents halogen, —CN, —NO₂, C₁₋₆alkyl,polyfluoroC₁₋₆alkyl, —OH or C₁₋₄alkoxy;

X represents O, S or N—R^(a) where R^(a) together with R¹ completes afused imidazole or 4,5-dihydroimidazole ring;

Y represents —CH₂—, —CH(OH)—, —CH(CH₃)—, —CH₂O—, —O— or —S;

R¹ represents H, C₁₋₆alkyl, C₃₋₈cycloalkyl, C₂₋₆alkyl, C₂₋₆alkynyl orpolyfluoroC₁₋₆alkyl, said alkyl, cycloalkyl, alkenyl and alkynyl groupsbeing optionally substituted by halogen, —CN, —NO₂, aryl, heteroaryl,—COR⁶, —CO₂R⁶, —CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷, —NR⁶SO₂R⁷, —SO₃R⁶,—SO₂N(R⁶)₂, —OR⁶, —SR⁶ or —N(R⁶)₂; or when X is N—R^(a), R¹ togetherwith R^(a) completes a fused imidazole or 4,5-dihydroimidazole ring;

R² and R^(2a) each represents hydrogen, or R² and R^(2a) togethercomplete a fused lactam ring of 4–7 members;

R³ represents aryl, heteroaryl, C₁₋₆alkyl, polyfluoroC₁₋₆alkyl,C₃₋₈cycloalkyl or C₃₋₈cycloalkyl₁₋₆alkyl;

each R⁶ independently represents H, polyfluoroC₁₋₆alkyl, or C₁₋₆alkylwhich is optionally substituted with halogen, —CN, —NO₂, —OH, —SH, —NH₂,phenyl, C₁₋₄alkoxy, C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino,—CO₂H, —CO₂C₁₋₄alkyl, —CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂; or twoR⁶ groups attached to a single nitrogen atom may complete a heterocyclicring of from 3 to 12 members including the said nitrogen, the remainingatoms being selected from C, N, O and S, and the ring optionally bearingup to 3 substituents independently selected from C₁₋₆alkyl,polyfluoroC₁₋₆alkyl, C₂₋₇acyl, —OH and —CONH₂;

R⁷ represents R⁶ that is other than H;

“aryl” refers to phenyl which is optionally fused to a 5–7 memberedsaturated or unsaturated ring which may be carbocyclic or may compriseup to 3 heteroatoms selected from nitrogen, oxygen and sulphur, andwhich may be oxo-substituted, said phenyl and optional fused ringtogether bearing 0–3 substituents independently selected from C₁₋₆alkyl[which is optionally substituted with halogen, —CN, —NO₂, —OH, —SH,—NH₂, C₁₋₄alkoxy, C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino,—CO₂H, —CO₂C₁₋₄alkyl, —CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂],polyfluoroC₁₋₆alkyl, halogen, —CN, —NO₂, heteroaryl, —COR⁶, —CO₂R⁶,—CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷, —NR⁶SO₂R⁷, —SO₃R⁶, —SO₂N(R⁶)₂, —OR⁶, —SR⁶and —N(R⁶)₂;

“heteroaryl” refers to a heteroaromatic ring of 5 or 6 members, at leastone member being nitrogen, oxygen or sulphur and the remainder carbon,said zing optionally being fused to a 5–7 membered saturated orunsaturated ring which may be carbocyclic or may comprise up to 3heteroatoms selected from nitrogen, oxygen and sulphur, and which may beoxo-substituted, the heteroaromatic ring and optional fused ringtogether bearing 0–3 substituents independently selected from C₁₋₆alkyl[which is optionally substituted with halogen, —CN, —NO₂, —OH, —SH,—NH₂, C₁₋₄alkoxy, C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino,—CO₂H, —CO₂C₁₋₄alkyl, —CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂],polyfluoroC₁₋₆alkyl, halogen, —CN, —NO₂, phenyl, —COR⁶, —CO₂R⁶,—CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷, —NR⁶SO₂R⁷, —SO₃R⁶, —SO₂N(R⁶)₂, —OR⁶, —SR⁶and —N(R⁶)₂;

or a pharmaceutically acceptable salt thereof

Where a variable occurs more than once in formula I or in a substituentthereof, the individual occurrences of that variable are independent ofeach other, unless otherwise specified.

As used herein, the expression “C_(1-x)alkyl” where x is an integergreater than 1 refers to straight-chained and branched alkyl groupswherein the number of constituent carbon atoms is in the range 1 to x.Particular alkyl groups are methyl, ethyl, n-propyl, isopropyl andt-butyl. Derived expressions such as “C₂₋₆alkenyl”, “hydroxyC₁₋₆alkyl”,“heteroarylC₁₋₆alkyl”, “C₂₋₆alkynyl” and “C₁₋₆alkoxy” are to beconstrued in an analogous manner.

The expression “polyfluoroC₁₋₆alkyl” as used herein refers to alkylgroups as defined above comprising at least one —CF₂— and/or —CF₃ group.

The expression “C₂₋₇acyl” as used herein refers to aromatic or linear,branched or cyclic aliphatic keto groups of up to 7 carbon atomsincluding the carbonyl group. Halogenated derivatives are encompassed.Examples include acetyl, trifluoroacetyl, benzoyl, n-propanoyl,isopropanoyl and cyclopentanoyl.

As used herein, the expression “C_(3-x)cycloalkyl” where x is an integergreater than 3 refers to nonaromatic hydrocarbon ring systems comprisingfrom 3 to x ring atoms. Where the specified number of ring atomspermits, the definition includes polycyclic systems, includingspirocyclic ortho-fused (including benzo-fused, provided attachment ofthe cycloalkyl group is via the non-aromatic ring) and bridged bicyclicsystems. “Spirocyclic” refers to a pair of rings having a single atom incommon. “Ortho-fused” refers to a pair of rings having two adjacentatoms in common. “Bridged bicyclic” refers to a pair of rings having atleast three adjacent atoms in common. Examples of cycloalkyl groupstherefore include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclohexenyl, indanyl, decalinyl, and bicyclo[2,2,1]hept-1-yl.

As used herein, the expression “heterocyclic ring” refers to monocyclicring systems comprising ring atoms selected from carbon, oxygen,nitrogen and sulphur, at least one ring atom being other than carbon.Examples include azetidine, pyrrolidine, piperidine, piperazine,morpholine and thiomorpholine.

As used herein, the expression “aryl” refers to phenyl which isoptionally fused to a 5–7 membered saturated or unsaturated ring whichmay be carbocyclic or may comprise up to 3 heteroatoms selected fromnitrogen, oxygen and sulphur, and which may be oxo-substituted, saidphenyl and optional fused ring together bearing 0–3 substituents asdescribed previously. The definition thus includes substituted andunsubstituted phenyl and naphthyl groups, and also groups comprising aphenyl ring which is fused to a saturated or unsaturated carbocyclic orheterocyclic ring, provided attachment of the aryl group is via thephenyl ring. The fused ring may be oxo-substituted, and hence may be acyclic lactone or lactam. Examples of aryl groups therefore also includemethylenedioxyphenyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl,benzofuranyl, indolyl and 2-oxoisoindolyl.

As used herein, the expression “heteroaryl” refers to a heteroaromaticring of 5 or 6 members, at least one member being nitrogen, oxygen orsulphur and the remainder carbon, said ring optionally being fused to a5–7 membered saturated or unsaturated ring which may be carbocyclic ormay comprise up to 3 heteroatoms selected from nitrogen, oxygen andsulphur, and which may be oxo-substituted, the heteroaromatic ring andoptional fused ring together bearing 0–3 substituents as describedpreviously. Generally, not more than 4, and preferably not more than 3atoms of the heteroaromatic ring are other than carbon. Where aheteroaromatic ring comprises two or more atoms which are not carbon,not more than one of said atoms may be other than nitrogen. Examples ofheteroaromatic rings include pyridine, pyridazine, pyrimidine, pyrazine,pyrrole, furan, thiophene, pyrazole, oxazole, isoxazole, thiazole,isothiazole, imidazole, oxadiazole, triazole, thiadiazole, tetrazole,1,2,4-triazine and 1,3,5-triazine. The optional fused ring may besaturated or unsaturated, including rings which are themselves(hetero)aromatic. Thus, for example, benzo-fused derivatives of theabove-listed heteroaromatic rings (where they are possible) are includedwithin the definition, provided attachment of the heteroaryl group isvia the heteroaromatic ring.

When a hydroxy substituent is present on a heteroaromatic ring andketo-enol tautomerism is possible, both tautomers are to be consideredas falling within the scope of the invention.

The term “halogen” as used herein includes fluorine, chlorine, bromineand iodine.

For use in medicine, the compounds of formula I may advantageously be inthe form of pharmaceutically acceptable salts. Other salts may, however,be useful in the preparation of the compounds of formula I or of theirpharmaceutically acceptable salts. Suitable pharmaceutically acceptablesalts of the compounds of this invention include acid addition saltswhich may, for example, be formed by mixing a solution of the compoundaccording to the invention with a solution of a pharmaceuticallyacceptable acid such as hydrochloric acid, sulphuric acid,methanesulphonic acid, fumaric acid, maleic acid, succinic acid, aceticacid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonicacid or phosphoric acid. Furthermore, where the compounds of theinvention carry an acidic moiety, suitable pharmaceutically acceptablesalts thereof may include alkali metal salts, e.g. sodium or potassiumsalts; alkaline earth metal salts, e.g. calcium or magnesium salts; andsalts formed with suitable organic ligands, e.g. quaternary ammoniumsalts.

Where the compounds according to the invention have at least oneasymmetric centre, they may accordingly exist as enantiomers. Where thecompounds according to the invention possess two or more asymmetriccentres, they may additionally exist as diastereoisomers. It is to beunderstood that all such isomers and mixtures thereof in any proportionare encompassed within the scope of the present invention. However, thestereochemistry at the position marked with an asterisk (*) in formula Iis preferably as shown in formula Ia:

In the compounds of formula I, n is preferably 0–2, most preferably 0 or1.

R^(x) is preferably halogen or C₁₋₆alkyl, most preferably halogen,especially chlorine. When n is 1, the substituent R^(x) is preferably inthe 7-position (i.e. para with respect to the nitrogen atom bonded toR¹).

X represents O, S or N—R^(a) where R^(a) combines with R¹ to complete afused imidazole or 4,5-dihydroimidazole ring. Typically X is O orN—R^(a), and preferably X is O.

Y represents —CH₂—, —CH(OH)—, —CH(CH₃)—, —CH₂O—, —O— or —S—; preferably—CH₂—, —CH(OH)—, —CH(CH₃)—, —CH₂O— or —O—.

Typically, R¹ represents H, polyfluoroC₁₋₆alkyl or C₁₋₆alkyl which isoptionally substituted with halogen, CN, aryl, heteroaryl, —CO₂R⁶,—CON(R⁶)₂, —OR⁶ or —N(R⁶)₂ where aryl, heteroaryl and R⁶ are as definedabove, or R¹ combines with X to complete a fused imidazole or4,5-dihydroimidazole ring. Preferably, R¹ represents H,polyfluoroC₁₋₆alkyl or C₁₋₄alkyl which is optionally substituted with—CN, —OH, azyl, heteroaryl, —CONH₂, C₁₋₆alkoxy or —N(R^(6a))₂ where eachR^(6a) independently represents H or C₁₋₆alkyl, or the two R^(6a) groupstogether complete a pyrrolidinyl, piperidinyl, piperazinyl ormorpholinyl ring, or R¹ combines with X to complete a fused imidazole or4,5-dihydroimidazole ring. Specific embodiments of R¹ include H, methyl,isopropyl, 2,2,2-trifluoroethyl, 4-methoxybenzyl,3-(morpholin-4-yl)propyl, 3-(pyrrolidin-1-yl)propyl, cyanomethyl,carbamoylmethyl, 5-chloro-1,2,3-thiadiazol-4-ylmethyl, 3-hydroxypropyland 3-dimethylaminopropyl. Most preferably, R¹ represents methyl or2,2,2-trifluoroethyl.

R² and R^(2a) are either both hydrogen, or together complete a fusedlactam ring of 4–7 members which may be fully saturated or may containunsaturation. Preferably, a fused lactam ring completed by R² and R^(2a)is 5- or 6-membered. In preferred embodiments, R² and R^(2a) are eitherboth hydrogen or together complete a fused pyrrolidinone or piperidinonering.

R³ may represent aryl, heteroalkyl, C₁₋₆alkyl, polyfluoroC₁₋₆alkyl,C₃₋₈-cycloalkyl or C₃₋₈cycloalkylC₁₋₆alkyl, but typically representspolyfluoroC₁₋₆alkyl (such as CF₃), aryl or heteroaryl. In particular, R³represents phenyl which optionally bears up to 3, but preferably notmore than 2, substituents independently selected from C₁₋₄alkyl,C₁₋₄alkoxy, trifluoromethyl and halogen atoms. Preferred embodiments ofR³ include phenyl, methylphenyl, methoxyphenyl,bis(trifluoromethyl)phenyl, chlorophenyl, fluorophenyl, dichlorophenyland difluorophenyl. Particularly preferred embodiments include2,4-dichlorophenyl, 2,4-difluorophenyl, 3,4-dichlorophenyl and3,4-difluorophenyl.

A subclass of the compounds of formula I is defined by formula II:

wherein:

R^(y), R^(z), R^(v) and R^(w) are independently H or halogen;

Y¹ is —CH₂—, —CH(OH)—, —CH(CH₃)—, —CH₂O— or —O—; and

R^(1a) is H, polyfluoroC₁₋₄alkyl, or C₁₋₄alkyl which is optionallysubstituted by —OH, —CN, carbamoyl or dimethylamino.

Another subclass of the compounds of formula I is defined by formulaIII:

wherein:

m is 1 or 2; and

R^(y), R^(z), R^(v), R^(w), Y¹ and R^(1a) are as defined above.

In the compounds of formulae II and III, preferably R^(z) is halogen andone of R^(v) and R^(w) is H while the other is halogen. Most preferably,R^(z) and R^(w) are both chlorine or both fluorine and R^(v) is H. R^(y)is preferably H or chlorine, most preferably H.

Examples of compounds in accordance with the invention include thosedisclosed in the Examples appended hereto, and pharmaceuticallyacceptable salts thereof.

The invention also provides pharmaceutical compositions comprising oneor more compounds of this invention and a pharmaceutically acceptablecarrier. Preferably these compositions are in unit dosage forms such astablets, pills, capsules, powders, granules, sterile parenteralsolutions or suspensions, metered aerosol or liquid sprays, drops,ampoules, transdermal patches, auto-injector devices or suppositories;for oral, parenteral, intranasal, sublingual or rectal administration,or for administration by inhalation or insufflation. For preparing solidcompositions such as tablets, the principal active ingredient is mixedwith a pharmaceutical carrier, e.g. conventional tableting ingredientssuch as corn starch, lactose, sucrose, sorbitol, talc, stearic acid,magnesium stearate, dicalcium phosphate or gums or surfactants such assorbitan monooleate, polyethylene glycol, and other pharmaceuticaldiluents, e.g. water, to form a solid preformulation compositioncontaining a homogeneous mixture of a compound of the present invention,or a pharmaceutically acceptable salt thereof. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulationcomposition is then subdivided into unit dosage forms of the typedescribed above containing from 0.1 to about 500 mg of the activeingredient of the present invention. Typical unit dosage forms containfrom 1 to 100 mg, for example 1, 2, 5, 10, 25, 50 or 100 mg, of theactive ingredient. The tablets or pills of the novel composition can becoated or otherwise compounded to provide a dosage form affording theadvantage of prolonged action. For example, the tablet or pill cancomprise an inner dosage and an outer dosage component, the latter beingin the form of an envelope over the former. The two components can beseparated by an enteric layer which serves to resist disintegration inthe stomach and permits the inner component to pass intact into theduodenum or to be delayed in release. A variety of materials can be usedfor such enteric layers or coatings, such materials including a numberof polymeric acids and mixtures of polymeric acids with such materialsas shellac, cetyl alcohol and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavoured syrups, aqueous or oilsuspensions, and flavoured emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles. Suitable dispersing or suspendingagents for aqueous suspensions include synthetic and natural gums suchas tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose,methylcellulose, poly(vinylpyrrolidone) or gelatin.

The compounds of the invention are potent inhibitors of γ-secretase,with the ability to arrest the production of β-amyloid peptide, andhence are useful in the treatment or prevention of diseases involvingthe deposition of β-amyloid.

Therefore, in a further aspect of the invention, there is provided theuse of a compound of formula I or a pharmaceutically acceptable saltthereof for the treatment or prevention of a condition associated withthe deposition of β-amyloid.

Preferably, the condition is a neurological disorder having associatedβ-amyloid deposition, such as Alzheimer's disease.

The present invention further provides the use of a compound of formulaI or a pharmaceutically acceptable salt thereof in the manufacture of amedicament for treating or preventing Alzheimer's disease.

Also disclosed is a method of treatment of a subject suffering from orprone to Alzheimer's disease which comprises administering to thatsubject an effective amount of a compound of formula I or apharmaceutically acceptable salt thereof.

For treating or preventing Alzheimer's Disease, a suitable dosage levelis about 0.01 to 250 mg/kg per day, preferably about 0.01 to 100 mg/kgper day, and especially about 0.01 to 5 mg/kg of body weight per day.The compounds may be administered on a regimen of 1 to 4 times per day.In some cases, however, dosage outside these limits may be used.

The compounds of formula I wherein X=O and R²=R^(2a)=H may be preparedby reaction of a compound of formula IV with a compound of Formula V:

where Z represents a protecting group such as benzyloxycarbonyl, andR^(x), n, R¹, R³ and Y have the same meanings as before. The processinvolves removal of the protecting group Z by treatment with acid (e.g.a solution of HBr in acetic acid), followed by coupling of the resultingprimary amine with carboxylic acid V. Any of the standard couplingmethods may be used, such as treatment with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC),hydroxybenzotriazole hydrate (HOBt) and triethylamine indichloromethane, or treatment withO-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU) and triethylamine in acetonitrile.

The protected amine IV may be prepared by a process comprising reactionof the benzodiazepinedione VI with Grignard reagent VII to form adductVIII:

conversion of VIII to the protected amine IX:

conversion of the oxazolidine group of IX to carboxylic acid to form X:

and conversion of the carboxylic acid group of X to carboxamide to formIV; wherein Boc represents t-butoxycarbonyl and Z, R^(x), n and R¹ havethe same meanings as before.

Diones of formula VI are available by the methods disclosed inWO97/49690, and Grignard reagent VII by the method described in J. Org.Chem., 1974, 39, 2790. Adduct VIII is formed by reaction of VI with VIIat −78° C. in THF, followed by treatment with HCl gas and saturatedNaHCO₃ solution. Conversion VIII to the protected amine IX involvesreaction of VIII with triisopropylbenzenesulfonyl azide in the presenceof strong base (e.g. potassium hexamethyldisilazide) in THF at −78° C.,followed by reduction of the resulting azide to the amine and additionof the protecting group Z. Any of the standard methods of reduction maybe used, such as hydrogenation over a Pd/C catalyst. Conversion of theoxazolidine group to carboxylic acid may be achieved by sequentialtreatment of IX with dilute HCl, acetyl chloride and dilute NaOH.Finally, the conversion of the carboxylic acid group to carboxamide maybe effected by conventional routes, e.g. treatment of X with ammoniumchloride, EDC, HOBt and triethylamine in DMF solution.

An alternative process for the preparation of protected amines IVcomprises reaction of a benzophenone XI with an N-protectedbenzotriazolylglycine XII to form the benzodiazepine derivative XIII:

optional N-alkylation of XIII to form the 1-substituted benzodiazepineXIV:

and replacement of the bromine atom of XIII or XIV with a carbamoylgroup to provide IV;wherein Bt represents benzotriazol-1-yl, R^(1b) represents R¹ that isother than H, and R^(x), n and Z have the same meanings as before.

Benzophenones XI may be prepared as described in J. Chem. Soc. PerkinTrans. I, 1995, 203–212, and glycine derivative XII as described in J.Org. Chem. 55, 2206 (1990), while the reaction to form XIII may becarried out as described in J. Org. Chem. 60, 730–4 (1995). N-alkylationof XIII may carried out by conventional means, e.g. reaction with sodiumhydride and R^(1b)-Hal in DMF, where Hal represents a leaving group suchas halide, especially iodide, and R^(1b) has the same meaning as before.Replacement of the bromine atom of XIV with a carbamoyl group may beachieved by reaction of XIV with carbon monoxide andhexamethyldisilazane in the presence of palladium (II) acetate,bis(diphenylphosphino)propane and a tertiary amine in DMF solution,followed by acid hydrolysis.

Compounds of formula I wherein X=O and R² and R^(2a) complete a fusedlactam zing may be prepared by reaction of the chlorides XV with theboronic acid derivatives XVI:

where p is 0, 1, 2 or 3, each R is independently H or C₁₋₆alkyl or thetwo R groups together complete a borolane ring, and R^(x), n, R¹, R³ andY have the same meanings as before. In particular, the R groups mayindividually represent H or together complete a4,4,5,5-tetramethyl-1,3,2-dioxaborolane ring. The reaction takes placein the presence of a Pd(0) catalyst in DMF solution at elevatedtemperature in a sealed vessel.

The chlorides XV are available by a process comprising coupling ofglycine derivative XII with aminobenzamide XVII to form amide XVIII:

cyclisation of XVIII to form benzodiazepinedione XIX:

coupling of XIX with carboxylic acid V to form amide XX:

and treatment of XX with phosphoryl chloride to form chloride XV; whereZ, Bt, R^(x), n, R¹, R³ and Y have the same meanings as before.

Coupling of XII with XVII is most readily achieved by conversion of thecarboxylic acid group of XII to the acid chloride (e.g. by treatmentwith oxalyl chloride) prior to reaction with the amino group of XVII.Cyclisation of XVIII is effected by heating in DMSO at about 180° C.Coupling of XIX with V may be carried out under the same conditions asthe reaction of IV with V. Conversion of XX to XV is effected byrefluxing briefly (approx. 10 minutes) in phosphoryl chloride.

An alternative synthesis of the chlorides XV, particularly suitable forembodiments wherein R¹ represents H or polyfluoroalkyl, comprisesring-opening of isatoic anhydride XXI with 2,4,6-trimethoxybenzylamineto form amide XXII:

reaction of XII with bromoacetyl bromide to form benzodiazepinedioneXXIII:

N-alkylation of XXIII to provide XXIV:

conversion of XXIV to the azide XXV and reduction thereof to the amineXXVI:

coupling of amine XXVI with carboxylic acid V followed by cleavage ofthe trimethoxybenzyl group to provide XX; and treatment of XX withphosphoryl chloride to provide chloride XV as described previously;where TMB represents 2,4,6-trimethoxybenzyl, and R^(x), n and R^(1b)have the same meanings as before.

Ring opening of XXI may be effected by refluxing withtrimethoxybenzylamine in a solvent such as ethyl acetate in the presenceof a tertiary amine. Cyclisation of XXI may be effected by treatmentwith bromoacetyl bromide at low temperature in the presence of alkali,and rafluxing of the resulting bromoacetalinide intermediate in sodiumisopropoxide solution. N-alkylation of XXIII may be effected by anyconventional means, but when R^(1b) represents 2,2,2-trifluoroethyl, aparticularly suitable process comprises treatment of XXII with2,2,2-trifluoroethyl iodide and caesium carbonate in DMF at atemperature of about 55° C. Conversion of XXIV to the azide XXV may becarried out by treatment with potassium t-butoxide and2,4,6-triisopropylbenzenesulfonyl azide in THF at −78° C., whilereduction of the azide to provide amine XXVI is possible by of theconventional routes, notably treatment with triphenylphosphine inaqueous THF. Coupling of the amine XXVI with carboxylic acid V ispossible by any of the conventional methods described previously, whilecleavage of the TMB group involves treatment with a mixture oftrifluoroacetic acid, water and dimethyl sulfide (approximately 90/5/5by volume).

It will be appreciated that a given compound in accordance with formulaI may be converted to another compound of formula I by the applicationof known synthetic techniques. For example, compounds of formula I inwhich X represents S may be prepared by treatment with Lawesson'sreagent of the corresponding compounds in which X represents O.Alternatively, and advantageously, this reaction may be carried out onthe synthetic precursors of such compounds, such as the compounds offormula IV. The reaction may be carried out as described in WO95/14693.The compounds of formula I, or the precursors thereof, wherein Xrepresents N—R^(a) may be prepared from the corresponding compounds inwhich X represents S using the methods disclosed in WO95/14693.

Similarly, a compound of formula I, or precursor thereof, in which R¹represents hydroxyalkyl may be converted to the mesylate by reactionwith methanesulfonyl chloride, and subjected to nucleophilicdisplacement with a primary or secondary amine to provide thecorresponding aminoalkyl derivative.

The starting materials V, VI, VII, XI, XII, XVI, XVII and XXI, wherethey are not commercially available, may be prepared by standardprocedures well known from the art, or by methods analogous to thosedescribed in detail hereinafter.

It will be appreciated that where more than one isomer can be obtainedfrom a reaction then the resulting mixture of isomers can be separatedby conventional means.

Where the above-described process for the preparation of the compoundsaccording to the invention gives rise to mixtures of stereoisomers,these isomers may be separated by conventional techniques such aspreparative chromatography. The novel compounds may be prepared inracemic form, or individual enantiomers may be prepared either byenantiospecific synthesis or by resolution. The novel compounds may, forexample, be resolved into their component enantiomers by standardtechniques such as preparative HPLC, or the formation of diastereomericpairs by salt formation with an optically active acid, such as(−)-di-p-toluoyl-d-tartaric acid and/or (+)di-p-toluoyl-l-tartaric acid,followed by fractional crystallization and regeneration of the freebase. The novel compounds may also be resolved by formation ofdiastereomeric esters or amides, followed by chromatographic separationand removal of the chiral auxiliary.

During any of the above synthetic sequences it may be necessary and/ordesirable to protect sensitive or reactive groups on any of themolecules concerned. This may be achieved by means of conventionalprotecting groups, such as those described in Protective Groups inOrganic Chemistry, ed. J. F. W. McOmie, Plenum Press, 1973; and T. W.Greene. & P. G. M. Wuts, Protective Groups in Organic Synthesis, JohnWiley & Sons, 1991. The protecting groups may be removed at a convenientsubsequent stage using methods known from the art.

Assays which can be used to determine the level of activity of compoundsof the present invention are disclosed in Biochemistry, 2000, 39(30),8698–8704.

The Examples of the present invention all had an ED₅₀ of less than 10μM, in preferred cases less than 1 μM, and in most preferred cases lessthan 100 nM in at least one of these assays.

EXAMPLES

The following schemes are representative of the methods used to preparethe compounds of the invention.Procedures

Scheme 1

Step 1A. To a stirred solution of2-(4-bromophenyl)-4,4-dimethyl-4,5-dihydrooxazole (J. Org. Chem. 1974,39, 2790) (9.15 g, 36.0 mmol.) in THF (100 ml) under nitrogen was addedmagnesium turnings (950 mg, 43.2 mmol.) and several crystals of iodine.The vigorously stirred mixture was gently warmed until the reaction hadinitiated. The mixture was allowed to self-reflux for 20 minutes andstirred a further 1 hour at room temperature. The resulting deep brownsolution was added via cannula to a −78° C. solution of tert-butyl1-methyl-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(WO 97/49690) (9.50 g, 32.8 mmol.) in THF (100 ml) and stirred at −78°C. for 20 minutes. The cooling bath was removed and the reaction stirredfor a further 2 hours after which time a saturated solution of NH₄Cl(100 ml) was added. The mixture was extracted into ethyl acetate (2×150ml) and the combined organics dried (MgSO₄), evaporated and purified bycolumn chromatography (SiO₂; ether) to afford the adduct 12.05 g, (79%)as an off-white solid. (¹H, CDCl₃) [exists as a ca. 4:1 mixture ofrotamers—data for major rotamer only reported] 8.03 (2H, d, J=7 Hz),7.79 (2H, d, J=8.5 Hz), 7.62 (1H, m), 7.47 (1H, m), 7.36 (1H, d, J=8Hz), 5.36 (1H, br s), 4.14 (2H, s), 3.81 (1H, dd, J=18, 6 Hz), 3.60 (1H,dd, J=18 Hz), 3.08 (3H, s,) 1.40 (9H, s) and 1.39 (6H, s).

Into a stirred solution of the Boc-protected amine (12.0 g, 26 mmol.) inethyl acetate (600 ml) cooled to −5° C. was bubbled HCl gas for 2.5hours. After this time the solvent was evaporated to give a solid whichwas redissolved in a mixture of THF (200 ml) and saturated aqueousNaHCO₃ (300 ml). The mixture was vigorously stirred for 1 hour andextracted into ethyl acetate (2×300 ml). Drying (MgSO₄) and evaporationafforded the product as a solid (8.9 g, 99%). (¹H, CDCl₃) 7.96 (2H, d,J=8 Hz), 7.65 (2H, d, J=8 Hz), 7.56 (1H, t, J=8 Hz), 7.36 (1H, d, J=8Hz), 7.25 (1H, d, J=8 Hz), 7.18 (1H, t, J=8 Hz), 4.83 (1H, d, J=10.5Hz), 4.13 (2H, s), 3.79 (1H, d, J=10.5 Hz), 3.43 (3H, s) and 1.40 (6H,s).

Step 1B. To a stirred solution of benzodiazepinone (10.6 g, 30 mmol.) inTHF (300 ml) at −78° C. was added potassium hexamethyldisilazide (0.5Msolution in toluene, 86 ml, 43 mmol.) portionwise over 15 minutes andthe mixture stirred for 10 minutes at −78° C. After this time,2,4,6-triisopropylbenzenesulfonyl azide (10.86 g, 35 mmol.) as asolution in THF (75 ml) was added via cannula and the reaction stirred afurther 10 minutes. A mixture of acetic acid (4 ml) and THF (75 ml) wasthen added, the cooling bath removed and the mixture stirred for 90minutes. Saturated NaHCO₃ solution (200 ml) was added and the mixtureextracted into ethyl acetate (3×150 ml). The combined organics weredried (MgSO₄) and evaporated to give a solid which was triturated withether to afford the desired azide as a colourless solid (9.1 g, 77%).(¹H, CDCl₃) 7.99 (2H, d, J=8.5 Hz), 7.72 (2H, d, J=8.5 Hz), 7.62 (1H, t,J=8 Hz), 7.40 (1H, d, J=8 Hz), 7.32 (1H, d, J=8 Hz), 7.25 (1H, t, J=8Hz), 4.56 (1H, s), 4.14 (2H, s), 3.49 (3H, s) and 1.40 (6H, s).

A solution of the azide (5.95 g, 15 mmol.) in ethanol (150 ml) wasdegassed with nitrogen bubbling for 10 minutes and then 5% palladium oncharcoal (100 mg) added and the mixture hydrogenated at 35 psi H₂ for 60minutes. The mixture was filtered through a pad of Celite washing wellwith ethanol and the combined organics evaporated to afford the amine(5.5 g 99%). (¹H, CDCl₃) 7.96 (2H, d, J=8 Hz), 7.66 (2H, d, J=8 Hz),7.58 (1H, t, J=8 Hz), 7.37 (1H, d, J=8 Hz), 7.27 (1H, d, J=8 Hz), 7.20(1H, t, J=8 Hz), 4.49 (1H, s), 4.13 (2H, s), 3.48 (3H, s) and 1.40 (6H,s).

To a stirred solution of the amine (5.6 g, 15.4 mmol.) and sodiumcarbonate (1.97 g, 18.6 mmol.) in a mixture of dioxan (200 ml) and water(100 ml) at 0° C. was added benzyl chloroformate (2.4 ml, 16.8 mmol.)dropwise. The mixture was stirred for 75 minutes at 0° C., quenched withsaturated ammonium chloride solution (200 ml) and extracted into ethylacetate (2×200 ml). The combined organics were dried (MgSO₄) andevaporated to afford the product as a foam (7.7 g, 99%). (1H, CDCl₃)7.95 (2H, d, J=8 Hz), 7.65–7.59 (3H, m), 7.39–7.23 (8H, m), 6.72 (1H, d,J=8 Hz), 5.32 (1H, d, J=8 Hz), 5.15 (2H, ABq), 4.14 (2H, s), 3.48 (3H,s) and 1.40 (6H, s).

Step 1C. The oxazoline (7.7 g, 15.5 mmol.) was dissolved in a mixture ofdioxan (50 ml) and 1M HCl (150 ml) and stirred at ambient temperaturefor 24 h. After this time, the mixture was cautiously basified withsodium carbonate solution and extracted into ethyl acetate (3×150 ml)and dichloromethane (2×100 ml). The combined organic extracts were dried(MgSO₄), evaporated and then redissolved in dichloromethane (100 ml).The solution was cooled to 0° C., triethylamine (1.4 ml, 10 mmol.) andacetyl chloride (0.66 ml, 9.2 mmol.) added and the mixture stirred atambient temperature for 1 hour. The solvent was evaporated and theresidue taken up in a mixture of THF (100 ml) and 1N NaOH (30 ml) andthe mixture stirred for a further 18 hours. After this time, thesolution was washed with ether (100 ml) and the aqueous layer acidifiedto pH 2 with 1N HCl and extracted into ethyl acetate (3×100 ml). Thecombined ethyl acetate layers were dried (MgSO₄) and evaporated toafford the product as an oil (3.5 g, 51%). (¹H, CDCl₃) 8.09 (2H, d,J=8.5 Hz) 7.75–7.59 (4H, m), 7.41–7.24 (7H, m), 6.74 (1H, d, J=8 Hz),5.34 (1H, d, J=8 Hz), 5.15 (2H, ABq) and 3.48 (3H, s).

Step 1D. To a stirred solution of carboxylic acid (4.25 g, 9.6 mmol.) inDMF (75 ml) was added ammonium chloride (5.0 g, 95 mmol.), EDC (2.21 g,11.5 mmol.), HOBt (1.56 g, 11.5 mmol.) and triethylamine (20 ml) and theMixture stirred at ambient temperature for 18 h. The solvent wasevaporated and the residue taken up in ethyl acetate (100 ml), washedwith 1N HCl (100 ml), saturated NaHCO₃ solution (100 ml) and water(3×100 ml). The organic layer was dried (MgSO₄) and evaporated to give ayellow powder which was triturated with ether to afford an off-whitepowder (2.7 g, 64%). (¹H, CDCl₃) 7.82 (2H, d, J=8.5 Hz), 7.75–7.59 (4H,m), 7.41–7.23 (7H, m) 6.72 (1H, d, J=8 Hz), 6.1 (1H, br s), 5.65 (1H,br, s), 5.33 (1H, d, J=8 Hz), 5.15 (2H, ABq) and 3.48 (3H, s).

To this benzyl carbamate (1.45 g, 3.3 mmol.) was added HBr (45% inacetic acid, 9 ml) and the mixture stirred at ambient temperature untilall the starting material had dissolved (35 minutes). The resultingbright orange solution was poured into ice cold ether (150 ml) andvigorously stirred for 10 minutes at 0° C. and filtered. The resultingpale yellow solid was partitioned between 4N NaOH (75 ml) anddichloromethane (100 ml), the layers separated and the aqueous layerextracted with further dichloromethane (3×100 ml) and 10% v/vmethanol/dichloromethane (2×100 ml). The combined organic layers weredried (MgSO₄) and evaporated to give a yellow semi-solid which wastriturated with ether to afford the product as an off-white powder (850mg, 84%). (¹H, CDCl₃) 7.83 (2H, d, J=8 Hz), 7.72 (2H, d, J=8 Hz), 7.60(1H, t, J=8 Hz), 7.38 (1H, d, J=8 Hz), 7.28 (1H, d, J=8 Hz), 7.22 (1H,t, J=8 Hz), 6.72 (1H, d, J=8 Hz), 6.15 (1H, br s), 5.65 (1H, br s), 4.49(1H, s) and 3.48 (3H, s).

Step 1E Representative Procedures.

(i), To a stirred solution of benzodiazepine amine (0.3 mmol.) indichloromethane or DMF (5 ml) under nitrogen was added the carboxylicacid (0.33 mmol.), EDC (0.33 mmol.), HOBt (0.33 mmol.) and triethylamine(0.6 mmol.) and the mixture stirred at ambient temperature for 12–24 h.The mixture was diluted with further dichloromethane (25 ml), washedsuccessively with 1N HCl (25 ml) [this washing omitted for productsbearing basic centres], 1N NaOOH (25 ml) and brine, dried (MgSO₄) andevaporated. The residue was purified by HPLC, column chromatography orpreparative thin layer chromatography on silica using an appropriateeluent.

(ii), To a stirred solution of benzodiazepine amine (0.3 mmol.) inacetonitrile (5 ml) under nitrogen was added the carboxylic acid (0.33mmol.), HBTU (0.33 mmol.) and triethylamine (0.6 mmol.) and the mixturestirred at ambient temperature for 12–24 h. The mixture was lyophilizedand the residue purified by HPLC using an appropriate eluent.

The product from Step 1D (designated A) could alternatively be preparedby the route shown in Scheme 2.

Scheme 2

Step 2A.[5-(4-bromophenyl)-2-oxo-2,3-dihydro-1H-benzo[e][1,4]-diazepin-3-yl]-carbamicacid benzyl ester.

2-Amino-4′-bromobenzophenone (J. Chem. Soc, Perkin Trans. 1, 1995,203–212) and the 2-(benzotriazol-1-yl)-N-(benzyloxycarbonyl)glycine (A.R. Katritzky et al, J. Org. Chem., 1990, 55, 2206) were reacted in ananalagous fashion to that described in J. Org. Chem. 1995, 60, 730–4 togive the title compound. ¹H-NMR (DMSO) 5.02–5.07 (3H, m), 7.25–7.67(14H, m), 8.45 (1H, d).

Step 2B.[5-(4-bromophenyl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[e][1,4]-diazepin-3-yl]-carbamicacid benzyl ester.

The product from Step 2A (8 g, 0.0172 moles) was dissolved in DMF (120ml) and treated with a 60% dispersion of sodium hydride in mineral oil(760 mg, 0.019 moles) followed by iodomethane (2.94 g, 0.021 moles) andallowed to stir at ambient temperature for 16 hours. The reaction wasquenched with water (100 ml) and extracted into ethyl acetate (2×100ml). The combined organic layers were washed with water (100 ml) andbrine (100 ml), dried (MgSO₄) and evaporated in vacuo. Purification bychromatography (SiO₂, 1% diethylether/dichloromethane) followed bytrituration with ether afforded the title compound (3.5 g, 43%). ¹H NMR(DMSO) 3.38 (3H, s), 5.06 (2H, s), 5.09 (1H, d, J=8.5 Hz), 7.34–7.68(13H, m), 8.50 (1H, d).

Step 2C.[5-(4-carbamoyl-phenyl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl]-carbamicacid benzyl ester.

A solution of the product from Step 2B (3.5 g, 0.0073 moles),1,3-bis(diphenylphosphino)propane (305 mg, 0.00073 moles),hexamethyldisilane (10.8 ml, 0.0146 moles), andN,N-diisopropylethylamine (2.5 ml, 0.0146) in DMF were degassed withnitrogen bubbling for ten minutes. Palladium (II) acetate (162 mg,0.00073 moles) was added and the mixture degassed for a further fiveminutes. Carbon monoxide gas was bubbled through the reaction mixturefor 5 minutes at room temperature and then for 6 hours at 1100° C. Afterthis time, the reaction mixture was cooled and partitioned betweendichloromethane (50 ml) and water (50 ml). The aqueous layer wasextracted with further dichloromethane (3×50 ml) and the combinedorganic layers washed with water (100 ml) and brine (100 ml), dried(MgSO₄) and evaporated in vacuo. The residue was taken up in a mixtureof THF (150 ml) and 2M HCl (30 ml) and stirred at ambient temperaturefor one hour. The THF was then evaporated in vacuo and the residuepartitioned between dichloromethane (50 ml) and 2M NaOH (50 ml). Theaqueous layer was extracted with dichloromethane (2×50 ml) and thecombined organic layers washed (H₂O, brine), dried (MgSO₄) andevaporated in vacuo. Purification by chromatography (SiO₂, 1%MeOH/CHCl₃) gave the title compound. ¹H NMR (DMSO) 3.31 (3H, s), 5.07(2H, s), 5.12 (1H, d), 7.30–7.80 (12H, m), 7.93 (2H, d, J=8.4 Hz), 8.08(1H, br s), 8.50 (1H, d). MS (ES+) MH⁺=443

Step 2D.4-(3-amino-1-methyl-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-5-yl)-benzamide.

To the product from Step 2C (400 mg, 0.9 mmol.) was added hydrogenbromide (45 wt % in acetic acid, 2 ml) and the mixture stirred untildissolution was complete (30 minutes). After this time, the orangesolution was poured into ice cold ether (20 ml) and vigorously stirredfor 10 minutes. The resulting precipitate was filtered and washed withcold ether to give the title compound (220 mg, 80%).

¹H NMR (CDCl₃) 3.08 (3H, s), 4.50 (1H, s), 5.70 (1H, v br s), 6.15 (1H,v br s) 7.20–7.42 (5H, m), 7.57–7.85 (5H, m). MS (ES+) MH⁺=309.

Scheme 3

Step 3A.Benzotriazol-2-yl-[(2-carbamoyl-phenyl)-methyl-carbamoyl]-methyl}-carbamicacid benzyl ester

A solution of 2-benzotriazol-1-yl)-N-(benzyloxycarbonyl)glycine (A. R.Katritzky et al, J. Org. Chem., 1990, 55, 2206) (50 g, 0.15 mol) in THF(300 ml) at 0 C was treated slowly with oxalyl chloride (2.0 M inCH₂Cl₂, 81 ml, 0.16 mol) and DMF (1 ml). The reaction mixture wasstirred at 0 C for 2 h, then treated with a solution of2-(methylamino)benzamide (23 g, 0.15 mol) and 4-methylmorpholine (38 ml,0.35 mol) in THF (100 ml). The reaction mixture was stirred overnight at40 C, then filtered. The residue was partitioned between water and warmethyl acetate. The aqueous layer was extracted three times with ethylacetate. The combined extracts were combined with the original filtrate,dried (MgSO₄), filtered and evaporated in vacuo. Trituration with ethylacetate gave the product as a white powder (23 g, 33%). The motherliquors were evaporated and purified by column chromatography to give afurther quantity of the product (21 g, 30%). (¹H NMR, DMSO) 9.3 (1H, d),8.8 (1H, d), 8.15–6.90 (15H, m), 4.92–4.75 (2H, m), 3.12 (3H, d, J=4.2Hz).

Step 3B.(1-Methyl-2,5-dioxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepin-3-yl)-carbamicacid benzyl ester

The product from Step 3A (23 g, 0.05 mol) was added to DMSO (500 ml) at180 C. The reaction mixture was stirred at 180 C for 20 min, cooled anddiluted with 1 M NaOH (aq) and ether. The aqueous phase was extractedwith ethyl acetate (five times) and the combined organic phases werewashed with brine, dried, filtered and evaporated. Purification bycolumn chromatography gave the product (5.8 g, 34%) as a yellow solid.(¹, HMR, DMSO) 9.30 (1H, d, J=4.0 Hz), 9.0 (3H, br s), 7.75–7.68 (2H,m), 7.56 (1H, d, J=8.1 Hz), 7.45–7.41 (1H, m), 5.20 (1H, d, J=4.2 Hz),3.4 (3H, s).

Step 3C.3-(3,4Dichloro-phenyl)-N-(1-methyl-2,5-dioxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide.

The product from Step 3B (3.27 g, 9.64 mmol) was dissolved in 48%HBr—AcOH and stirred for 35 min. The reaction mixture was poured into alarge volume of ice-cold ether. The resulting precipitate was collectedby filtration, washed with ether and dried in vacuo. The product wasobtained as a white solid (2.65 g, 96%). (¹H NMR, DMSO) 8.73 (1H, br d,J=3.6 Hz), 7.74–7.32 (10H, m), 5.21 (1H, dd, J=4.6, 7.8 Hz), 5.06 (2H,s), 3.31 (3H, s). MH+=340, MNa+=352.

This product was coupled to 3-(3,4-dichlorophenyl)propionic acid understandard conditions to yield the desired product. 1H NMR (DMSO) 8.73(1H, d), 8.58 (1H, d) 7.74–7.31 (7H, m). 5.37 (1H, dd), 3.336 (3H, s)2.93–2.82 (2H, m), 2.65–2.51 (2H, m). m/z=407 (C₁₉H₁₇N₃Cl₂O₃+H⁺).

Step 3D.N-(5-Chloro-1-methyl-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-3-(2,4-dichloro-phenyl)-propionamide.

The product from step 3C (1 g, 2.46 mmol) was finely ground, suspendedin POCl₃ (ca 20 ml) and placed in an oil bath at 95 C. Afterapproximately 10 minutes, the reaction mixture was cooled, diluted withethyl acetate and added slowly to an ice-cold solution of NaHCO₃containing ice. The mixture was shaken vigorously. The aqueous layer wasseparated and the organic layer was washed thoroughly with NaHCO₃(twice), brine, dried (MgSO₄), filtered and evaporated in vacuo to givean orange solid. Trituration with ether gave the imidoyl chloride (0.7g, 67%). ¹H NMR (DMSO) 9.24 (1H, d, J=8.1), 7.87–7.74 (2H, m), 7.62–7.55(2H, m), 7.48–7.33 (3H, m), 5.24 (1H, d, J 8.1), 3.37 (3H, s), 2.90–2.86(2H, m), 2.61–2.55 (2H, m). m/z=424–428 (cluster) (C₁₉H₁₆N₃Cl₃O₂+H⁺).

Step 3E Representative Procedure

The product from step 3D (100 mg, 0.23 mmoles), tripotassium phosphate(84 mg, 0.4 mmoles),2-(1-oxo-2,3-dihydro-1H-isoindol-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxa-borolane(91 mg, 0.35 mmoles) and DMF (4 ml) in a thick-walled flask weredegassed with nitrogen. Pd(PPh₃)₄ was added and the vessel sealed andheated at 90° C. for 2 hours. The mixture was cooled and taken up inwater/ethyl acetate. The organic layer was washed (water, brine), dried(MgSO₄) and evaporated in vacuo. Purification by flash silica columneluting with ethyl acetate gave the title compound.

Abbreviations:

-   EDC—1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-   HOBt—Hydroxybenzotriazole hydrate-   HBTU—O-Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate

Example 1(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-(3-[morpholin-4-yl]propyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(3-(tert-butyldiphenylsilyloxy)propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690). During Step 1C, the silylprotecting group was cleaved and the resulting primary hydroxyltransformed to the morpholin-1-yl function by way of the mesylate usingstandard methods. Step 1E was carried out using3-(2,4-dichlorophenyl)propionic acid.

(¹H, CDCl₃) 1.64 (1H, m), 1.69 (1H, m), 2.15 (4H, m), 2.26 (2H, m), 2.70(2H, m), 3.09 (2H, dd, J=6.8, 6.8), 3.60 (4H, m), 3.79 (1H, m), 4.40(1H, m), 5.51 (2H, d, J=7.2), 5.82 (1H, brs), 6.21 (1H, brs), 7.17 (1H,m) 7.27 (3H, m), 7.38 (2H, m), 7.47 (1H, m), 7.61 (1H, m), 7.66 (2H, d,J=7.5), 7.83 (2H, d, J=7.5). MS(CI+): MH+=622

Example 2(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-2-oxo-1-(3-[pyrrolidin-1-yl]propyl)-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamidehydrochloride

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(3-(tert-butyldiphenylsilyloxy)-propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690). During Step 1C, the silylprotecting group was cleaved and the resulting primary hydroxyltransformed to the pyrrolidin-1-yl-function by way of the mesylate usingstandard methods. Step 1E was carried out using3-(2,4-dichlorophenyl)propionic acid.

(1H, DMSO) 1.82 (4H, m), 2.62–2.79 (4H, m), 2.90 (2H, m), 3.24 (1H, m),3.36 (1H, m), 3.59 (4H, m), 3.91 (1H, m), 4.23 (1H, m), 5.32 (1H, d,J=7.8), 7.37 (4H, m), 7.49 (1H, brs), 7.58 (1H, d, J=1.9), 7.66 (2H, d,J=8.2), 7.76 (2H, d, J=3.6), 7.96 (2H, d, J=8.2), 8.11 (1H, brs), 9.23(1H, d, J=7.8), 10.31 (1H, brs). MS(CI+): MH+=606

Example 3(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-[3-(dimethylamino)propyl]-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamidehydrochloride

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(3-(tert-butyldiphenylsilyloxy)-propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690). During Step 1C, the silylprotecting group was cleaved and the resulting primary hydroxyltransformed to the dimethylamino function by way of the mesylate usingstandard methods. Step 1E was carried out using3-(2,4-dichlorophenyl)-propionic acid.

(1H, DMSO) 1.88 (2H, m), 2.55 (3H, d, J=4.2), 2.59 (3H, d, J=4.2), 2.64(2H, m), 2.82 (1H, m), 2.92 (3H, m), 3.89 (1H, m), 4.22 (1H, m), 5.33(1H, d, J=7.7), 7.13 (1H, m), 7.24 (1H, m), 7.32 (2H, m), 7.37 (2H, m),7.51 (1H, d, J=7.7), 10.35 (1H, brs). MS(CI+): MH+=580

Example 4(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-(4-methoxybenzyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(4-methoxybenzyl)-propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690) and Step 1E was carried out using3-(2,4-dichlorophenyl)-propionic acid.

(¹H, CDCl₃) 2.71 (2H, m), 3.11 (2H, m), 3.69 (3H, s), 4.69 (1H, d,J=14.9 Hz), 5.59 (1H, d, J=14.9 Hz), 5.59 (1H, d, J=8.0 Hz), 5.65 (1H,broad s), 6.12 (1H, broad s), 6.61 (2H, d, J=8.7 Hz), 6.89 (2H, d, J=8.7Hz), 7.13–7.25 (4H, m), 7.34–7.40 (4H, m), 7.46–7.56 (2H, m), 7.76 (2H,d, J=8.4 Hz); MS(ES+), MH⁺:615

Example 5(±)-4-(3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl)benzamide

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(4-methoxybenzyl)-propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690). Step 1E was carried out using3(2,4-dichlorophenyl)-propionic acid and the 4methoxybenzyl grouppresent in the product of Step 1E was removed using ceric ammoniumnitrate under standard conditions.

(¹H, CDCl₃) 2.71 (2H, m), 3.09 (2H, m), 5.23 (1H, d, J=7.8), 6.23 (1H,brs), 6.41 (1H, brs), 7.18 (5H, m), 7.35 (1H, d, J=1.9), 7.42 (1H, m),7.50 (3H, m), 7.76 (2H, d, J=8.2), 9.3 (1H, s). MS(CI+): MH+=495

Example 6(±)-4-[3-{[3-(2,4-chlorophenyl)propanoyl]amino}-1-(3-hydroxypropyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by modification of the route shown in Scheme 1. The startingmaterial for Step 1A was tert-butyl1-(3-(tert-butyldiphenylsilyloxy)-propyl)-2,5-dioxo-1,2,3,5-tetrahydro-4H-1,4-benzodiazepine-4-carboxylate(prepared by analogy to WO 97/49690).

(¹H, CDCl₃) 1.6 (1H, m), 1.8 (1H, m), 2.71 (2H, ddd, J=2.3, 7.6, 7.6),3.09 (2H, dd, J=7.6, 7.6), 3.32 (1H, m), 3.38 (1H, m), 3.84 (1H, m),4.51 (1H, m), 5.53 (1H, d, J=8), 5.60 (1H, brs), 6.0 (1H, brs), 6.45(1H, brs), 7.16 (1H, m), 7.25 (3H, m), 7.37 (1H, d, J=2), 7.49–7.63 (5H,m), 7.82 (2H, d, J=8.2). MS(CI+): MH+=553

Example 7(±)-3-(2,4-Dichlorophenyl)-1-N-[2-oxo-2,3-dihydro-1H-isoindol-5-yl)-1-methyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide

Prepared by the route shown in Scheme 3. Step 3E was carried out using2-(1-oxo-2,3-dihydro-1H-isoindol-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxa-borolane.

(¹H, CDCl₃) 9.26 (1H, d, J=8.1), 8.70 (1H, s), 7.75–7.34 (10H, m), 5.34(1H, d, J=8.1), 4.43 (1H, d, J=12.0), 4.40 (1H, d, J=12.0), 3.39 (3H,s), 2.94–2.90 (2H, m), 2.65–2.61 (2H, m), MS (Electrospray): MH+=520

Example 8(±)-3-(3,4-Dichlorophenyl)-1-N-[2-oxo-5-(1-oxo-1,2,3,4-tetrahydro-6-isoquinolinyl)-1-methyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide

Prepared by the route shown in Scheme 3. Step 3E was carried out using2-(1-oxo-1,2,3,4-tetrahydro-6-isoquinolinyl)-4,4,5,5-tetramethyl-1,3,2-dioxa-borolane.

(¹H, CDCl₃) 9.27 (1H, d, J=8.1), 8.03 (1H, s), 7.92 (1H, d, J=8.0),7.77–7.34 (9H, m), 5.32 (1H, d, J=8.1), 3.40–3.35 (5H, m), 2.94–2.91(4H, m), 2.67–2.62 (2H, m), MS (Electrospray): MH+=534

Example 94-[3-{[(3S)-3-phenylbutyryl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide(1:1 Mixture of Diastereomers)

Prepared by reaction of amine A and (S)-3-phenylbutyric acid using theprocedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 1.36 (3H, d, J=6.9 Hz, single diast.), 1.38 (3H, d, J=7.0Hz, single diast.), 2.61 (1H, m), 2.69 (1H, m), 3.37 (1H, m) 3.46 (3H,s, single diast.), 3.47 (3H, s, single diast.), 5.48 (1H, d, J=7.9 Hz,single diast.), 5.51 (1H, d, J=8.1 Hz, single diast.), 5.63 (1H, broads), 6.12 (1H, broad s), 7.27 (2H, m), 7.29 (1H, m), 7.31 (2H, m), 7.33(1H, m), 7.37 (2H, m), 7.40 (1H, m), 7.60 (3H, m), 7.82 (2H, m);MS(ES+), MH⁺:455

Example 10(±)-4-[3-{[3-(3,4-difluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3,4-diflurophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.67 (2H, m), 2.98 (2H, m), 3.48 (3H, s), 5.52 (1H, d, J=8.0Hz), 5.61 (1H, broad s), 6.17 (1H, broad s), 6.93 (1H, m), 7.08 (2H, m),7.28 (2H, m), 7.41 (2H, m) 7.61 (1H, m) 7.67 (2H, m), 7.82 (2H, m)

Example 11(±)-4-[3-{[3-(3-chlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3-chlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.68 (2H, m), 2.98 (2H, m), 3.48 (3H, s), 5.53 (1H, d, J=7.9Hz), 5.77 (1H, broad s), 6.21 (1H, broad s), 7.12 (1H, m), 7.17–7.29(4H, m), 7.39 (3H, m), 7.59 (1H, m), 7.67 (2H, d, J=8.2 Hz), 7.83 (2H,d, J=8.2 Hz)

Example 12(±)-4-[3-{[3-(4-fluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(4-fluorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.68 (2H, m), 2.99 (2H, m), 3.48 (1H, s), 5.53 (1H, d, J=8.0Hz), 5.68 (1H, broad s), 6.12 (1H, broad s), 6.97 (2H, m), 7.19 (1H, m),7.32 (1H, m), 7.39–7.45 (4H, m), 7.62 (1H, m), 7.68 (2H, m), 7.84 (2H,m); MS(ES+), MH⁺:459

Example 13(±)-4-[3-{[3-(4-chlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(4-chlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.71 (2H, m), 3.03 (2H, m), 3.48 (3H, s), 5.54 (1H, d, J=8.0Hz), 5.68 (1H, broad s), 6.16 (1H, broad s), 7.18 (2H, d, J=8.4 Hz),7.25 (1H, m), 7.32 (1H, m), 7.39 (2H, m), 7.40 (2H, m), 7.59 (1H, m),7.67 (2H, d, J=8.4 Hz), 7.83 (2H, d, J=8.4 Hz); MS(ES+), MH⁺:475

Example 14(±)-4-[3-{[3-phenylpropanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-phenylpropionic acid using theprocedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.68 (2H, m), 3.00 (2H, t, J=7.7 Hz), 3.48 (3H, s), 5.52(1H, d, J=8.0 Hz), 5.65 (1H, broad s), 6.12 (1H, broad s), 7.19–7.41(8H, m), 7.45 (1H, m), 7.61 (1H, m), 7.67 (2H, d, J=8.4 Hz), 7.83 (2H,d, J=8.4 Hz);

MS(ES+), MH⁺:441

Example 15(±)-4-[3-{[3-(3,5-bis(trifluoromethyl)phenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and3-(3,5-bis(trifluoromethyl)phenyl)propionic acid using the procedure ofStep 1E shown in Scheme 1.

(¹H, CDCl₃) 2.75 (2H, m), 3.16 (2H, m), 3.48 (3H, s), 5.51 (1H, d, J=8.0Hz), 5.65 (1H, broad s), 6.10 (1H, broad s), 7.24–7.42 (4H, m),7.60–7.64 (6H, m), 7.82 (2H, d, J=8.4 Hz); MS(ES+), MH⁺:577

Example 16(±)-4-[3-{[3-(2,3-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(2,3-dichlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.72 (2H, m), 3.16 (2H, t, J=7.8 Hz), 3.48 (3H, s), 5.53(1H, d, J=7.9 Hz), 5.90 (1H, broad s), 6.25 (1H, broad s), 7.10–7.46(7H, m), 7.61 (1H, m), 7.66 (2H, d, J=8.3 Hz), 7.82 (2H, d, J=8.3 Hz);MS(ES+), MH⁺:509

Example 17(±)-4-[3-{[3-(3,4-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3,4-dichlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.68 (2H, m), 2.98 (2H, m), 3.48 (3H, s), 5.51 (1H, d, J=8.0Hz), 5.80 (1H, broad s), 6.15 (1H, broad s), 7.08 (1H, d, J=8.2 Hz),7.24–7.41 (6H, m), 7.60 (1H, m), 7.66 (2H, d, J=8.2 Hz), 7.83 (2H, d,J=8.2 Hz); MS(ES+), MH⁺:509

Example 18 (±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(2,4-dichlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.70 (2H, m), 3.11 (2H, m), 3.48 (3H, s), 5.52 (1H, d, J=8.0Hz), 5.65 (1H, broad s), 6.05 (1H, broad s), 7.16 (1H, dd, J=8.2, 2.0Hz), 7.24–7.41 (6H, m), 7.60 (1H, m), 7.67 (2H, d, J=8.2 Hz), 7.83 (2H,d, J=8.2 Hz); MS(ES+), MH⁺:509

Example 19(±)-4-[3-{[3-(3,5-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3,5-dichlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H; CDCl₃), 2.69 (2H, m), 2.97 (2H, m), 3.48 (3H, s), 5.52 (1H, d,J=8.0 Hz), 6.20 (2H, broad s), 7.12 (2H, m), 7.20–7.30 (3H, m), 7.40(1H, d, J=8.3 Hz), 7.48 (1H, d, J=8 Hz), 7.61 (1H, m), 7.66 (2H, d,J=8.2 Hz), 7.83 (2H, d, J=8.2 Hz); MS(ES+), MH⁺:509

Example 20(±)-4-[3-{[3-(3-methoxyphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3-methoxyphenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.70 (2H, m), 3.00 (2H, m), 3.48 (3H, s), 3.79 (3H, s), 5.54(1H, d, J=8.0 Hz), 5.75 (1H, broad s), 6.20 (1H, broad s), 6.74–6.85(3H, m), 7.19–7.41 (5H, m), 7.60 (1H, m), 7.67 (2H, d, J=8.2 Hz), 7.82(2H, d, J=8.2 Hz); MS(ES+), MH⁺:471

Example 21(±)-4-[3-{[3-(4-methoxyphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(4-methoxyphenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

MS(ES+) MH⁺:471

Example 22(±)-4-[3-{[3-(4-methylphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(4-methylphenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.33 (3H, s), 2.68 (2H, m), 2.99 (2H, m), 3.48 (3H, s), 5.54(1H, d, J=8.0 Hz), 5.73 (1H, broad s), 6.17 (1H, broad s), 7.10–7.15(4H, m), 7.24–7.42 (4H, m), 7.60 (1H, m), 7.68 (2H, d, J=8.4 Hz), 7.83(2H, d, J=8.4 Hz); MS(ES+), MH⁺:455

Example 23(±)-4-[3-{(2,4-dichlorophenoxyacetyl)amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 2,4-dichlorophenoxyacetic acid usingthe procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 3.51 (3H, s), 4.63 (2H, Abq, J=36.4, 14.4 Hz), 5.59 (1H, d,J=7.8 Hz), 5.70 (1H, broad s), 6.12 (1H, broad s), 6.90 (1H, d, J=8.8Hz), 7.22–7.45 (5H, m), 7.63 (1H, m), 7.70 (2H, d, J=8.4 Hz) 7.84 (2H,d, J=8.4 Hz) 8.67 (1H, d, J=7.71 Hz); MS(ES+), MH⁺:511

Example 24(±)-4-[3-{[3-(3,5-difluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(3,5-difluorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.69 (2H, m), 3.01 (2H, m), 3.48 (3H, s), 5.54 (1H, d, J=7.8Hz), 5.92 (1H, broad s), 6.28 (1H, broad s), 6.64 (1H, m), 6.77 (2H, d,J=6.3 Hz), 7.25–7.32 (2H, m), 7.41 (1H, d, J=8.3 Hz), 7.47 (1H, d, J=7.5Hz), 7.63 (1H, m), 7.67 (2E, d, J=8.0 Hz), 7.83 (2H, d, J=8.0 Hz);MS(ES+), MH⁺:477

Example 25(±)-4-[3-{[3-(2,5-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(2,5-dichlorophenyl)propionic acidusing the procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 2.70 (2H, m), 3.09 (2H, m), 3.48 (3H, s), 5.53 (1H, d, J=8.0Hz), 5.76 (1H, broad s), 6.19 (1H, broad s), 7.14 (1H, dd, J=8.5, 2.5Hz), 7.24–7.31 (4H, m), 7.40 (2H, dd, J=8.5, 2.3 Hz), 7.61 (1H, m), 7.68(2H, d, J=8.2 Hz), 7.83 (2H, d, J=8.2 Hz); MS(ES+), MH⁺:509

Example 26(±)-4-[3-{[3-(2,4-dichlorophenoxy)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3-(2,4-dichlorophenoxy)propionicacid using the procedure of Step 1E shown in Scheme 1.

(¹, (CDCl₃) 2.92 (2H, t, J=5.8 Hz), 3.48 (3H, s), 4.36 (2H, t, J=5.8Hz), 5.55 (1H, d, J=7.8 Hz), 5.72 (1H, broad s), 6.18 (1H, broad s),6.91 (1H, d, J=8.8 Hz), 7.16 (1H, dd, J=8.7, 2.4 Hz), 7.23–7.31 (3H, m),7.35 (1H, d, J=2.6 Hz), 7.40 (2H, dd, J=8.1, 0.5 Hz), 7.61 (1H, m), 7.68(2H, d, J=8.2 Hz), 7.83 (2H, d, J=8.2 Hz); MS(ES+), MH⁺:525

Example 27(±)-4-[3-{[2-(3,4-dichlorophenoxy)acetyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-4-benzodiazepin-5-yl]benzamide

Prepared by reaction of amine A and 3,4-dichlorophenoxyacetic acid usingthe procedure of Step 1E shown in Scheme 1.

(¹H, CDCl₃) 3.48 (3H, s), 4.59 (2H, dd, J=24.8, 14.7 Hz), 5.59 (1H, d,J=8.0 Hz), 5.77 (1H, broad s), 6.19 (1H, broad s), 6.89 (1H, dd, J=8.9,3.0 Hz), 7.13 (1H, d, J=3.0 Hz), 7.27–7.44 (4H, m), 7.63 (1H, m), 7.69(2H, d, J=8.3 Hz), 7.83 (2H, d, J=8.3 Hz), 8.36 (1H, d, J=8.0 Hz);MS(ES+), MH⁺:511

1. A compound of formula I:

wherein n is 0–3; each R^(x) independently represents halogen, —CN,—NO₂, C₁₋₆alkyl, polyfluoroC₁₋₆alkyl, —OH or C₁₋₄alkoxy; X represents O,S or N—R^(a) where R^(a) together with R¹ completes a fused imidazole or4,5-dihydroimidazole ring; Y represents —CH₂—, —CH(OH)—, —CH(CH₃)—,—CH₂O—, —O— or —S; R¹ represents H, C₁₋₆alkyl, C₃₋₈cycloalkyl,C₂₋₆alkenyl, C₂₋₆alkynyl or polyfluoroC₁₋₆alkyl, said alkyl, cycloalkyl,alkenyl and alkynyl groups being optionally substituted by halogen, —CN,—NO₂, aryl, heteroaryl, —COR⁶, —CO₂R⁶, —CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷,—NR⁶SO₂R⁷, —SO₃R⁶, —SO₂N(R⁶)₂, —OR⁶, —SR⁶ or —N(R⁶)₂; or when X isN—R^(a), R¹ together with R^(a) completes a fused imidazole or4,5-dihydroimidazole ring; R² and R^(2a) each represents hydrogen, or R²and R^(2a) together complete a fused lactam ring of 4–7 members; R³represents aryl, heteroaryl, C₁₋₆alkyl, polyfluoroC₁₋₆alkyl,C₃₋₈cycloalkyl or C₃₋₈cycloalkylC₁₋₆alkyl; each R⁶ independentlyrepresents H, polyfluoroC₁₋₆alkyl, or C₁₋₆alkyl which is optionallysubstituted with halogen, —CN, —NO₂, —OH, —SH, —NH₂, phenyl, C₁₋₄alkoxy,C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino, —CO₂H, —CO₂C₁₋₄alkyl,—CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂; or two R⁶ groups attached toa single nitrogen atom may complete a heterocyclic ring of from 3 to 12members including the said nitrogen, the remaining atoms being selectedfrom C, N, O and S, and the ring optionally bearing up to 3 substituentsindependently selected from C₁₋₆alkyl, polyfluoroC₁₋₆alkyl, C₂₋₇acyl,—OH and —CONH₂; R⁷ represents R⁶ that is other than H; “aryl” refers tophenyl which is optionally fused to a 5–7 membered saturated orunsaturated ring which may be carbocyclic or may comprise up to 3heteroatoms selected from nitrogen, oxygen and sulphur, and which may beoxo-substituted, said phenyl and optional fused ring together bearing0–3 substituents independently selected from C₁₋₆alkyl [which isoptionally substituted with halogen, —CN, —NO₂, —OH, —SH, —NH₂,C₁₋₄alkoxy, C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino, —CO₂H,—CO₂C₁₋₄alkyl, —CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂],polyfluoroC₁₋₆alkyl, halogen, —CN, —NO₂, heteroaryl, —COR⁶, —CO₂R⁶,—CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷, —NR⁶SO₂R⁷, —SO₃R⁶, —SO₂N(R⁶)₂, —OR⁶, —SR⁶and —N(R⁶)₂; “heteroaryl” refers to a heteroaromatic ring of 5 or 6members, at least one member being nitrogen, oxygen or sulphur and theremainder carbon, said ring optionally being fused to a 5–7 memberedsaturated or unsaturated ring which may be carbocyclic or may compriseup to 3 heteroatoms selected from nitrogen, oxygen and sulphur, andwhich may be oxo-substituted heteroaromatic ring and optional fused ringtogether bearing 0–3 substituents independently selected from C₁₋₆alkyl[which is optionally substituted with halogen, —CN, —NO₂, —OH, —SH,—NH₂, C₁₋₄alkoxy, C₁₋₄alkylthio, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino,—CO₂H, —CO₂C₁₋₄alkyl, —CONH₂, —CONHC₁₋₄alkyl or —CON(C₁₋₄alkyl)₂],polyfluoroC₁₋₆alkyl, halogen, —CN, —NO₂, phenyl, —COR⁶, —CO₂R⁶,—CON(R⁶)₂, —OCOR⁷, —NR⁶COR⁷, —NR⁶SO₂R⁷, —SO₃R⁶, —SO₂N(R⁶)₂, —OR⁶, SR⁶and —N(R⁶)₂; or a pharmaceutically acceptable salt thereof.
 2. Acompound according to claim 1 wherein the stereochemistry at theposition marked with an asterisk (*) in formula I is as shown in formulaIa:


3. A compound according to claim 1 of formula II:

wherein: R^(y), R^(z), R^(v) and R^(w) are independently H or halogen;Y¹ is —CH₂, —CH(OH)—, —CH(CH₃)—, —CH₂O— or —O—; and R^(1a) is H,polyfluoroC₁₋₄alkyl, or C₋₄alkyl which is optionally substituted by —OH,—CN, carbamoyl or dimethylamino; or a pharmaceutically acceptable saltthereof.
 4. A compound according to claim 1 of formula III:

wherein: m is 1 or 2; R^(y), R^(z), R^(v) and R^(w) are independently Hor halogen; Y¹ is —CH₂₋, —CH(OH)—, —CH(CH₃)—, —CH₂O— or —O—; and R^(1a)is H, polyfluoroC₁₋₄alkyl, or C₁₋₄alkyl which is optionally substitutedby —OH, —CN, carbamoyl or dimethylamino; or a pharmaceuticallyacceptable salt thereof.
 5. A compound according to claim 3 or claim 4wherein R^(z), halogen and one of R^(v) and R^(w) is H while the otheris halogen.
 6. A compound according to claim 1 selected from:(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-(3-[morpholin-4-yl]propyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-2-oxo-1-(3-[pyrrolidin-1-y]propyl)-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamidehydrochloride;(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-[3-(dimethylamino)propyl]-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamidehydrochloride;(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-(4-methoxybenzyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-(3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl)benzamide;(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-(3-hydroxypropyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-3-(2,4-dichlorophenyl)-1-N-[2-oxo-5-(1-oxo-2,3-dihydro-1H-isoindol-5-yl)-1-methyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide;(±)-3-(3,4-dichlorophenyl)-1-N-[2-oxo-5-(1-oxo-1,2,3,4-tetrahydro-6-isoquinolinyl)-1-methyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide;4-[3-{[(3S)-3-phenylbutyryl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3,4-difluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3-chlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(4-fluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(4-chlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-phenylpropanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3,5-bis(trifluoromethyl)phenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(2,3-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3,4-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(2,4-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3,5-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3-methoxyphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(4-methoxyphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(4-methylphenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{(2,4-dichlorophenoxy)acetyl)amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(3,5-difluorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(2,5-dichlorophenyl)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[3-(2,4-dichlorophenoxy)propanoyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide;(±)-4-[3-{[2-(3,4-dichlorophenoxy)acetyl]amino}-1-methyl-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-5-yl]benzamide.7. A pharmaceutical composition comprising one or more compoundsaccording to claim 1 and a pharmaceutically acceptable carrier.
 8. Amethod of treatment of a subject suffering from Alzheimer's diseasewhich comprises administering to that subject an effective amount of acompound according to claim 1.